阻塞性黄疸肾脏系膜细胞内信号传导的机制及其意义

The Mechanism and Significance of Cellular Signal Transduction in Kidney Glomerular Mesangial Cells of Rat with Obstructive Jaundice

目的了解神经酰胺介导的肾脏损伤中细胞内信号传导的机制以及在肾损害中的作用。方法将大鼠分为正常对照组、假手术组、胆总管结扎15 d和21 d组,每组15只大鼠。胆总管结扎组建立阻塞性黄疸大鼠模型(胆总管结扎),取肾脏行系膜细胞原代培养,分别给予不同的干预措施后,胆碱比色法测定细胞膜磷脂酶D(PC-PLD)活力,流式细胞仪分析细胞周期亚G1峰。结果使用肿瘤坏死因子-α(TNF-α)和外源性神经酰胺C2-C eram ide可使肾系膜细胞PLD活力下降(P<0.05),诱导肾脏系膜细胞凋亡,给予神经酰胺抑制剂可使系膜细胞凋亡率下降,PLD活性增加。结论TNF-α与膜上受体结合后可引起胞内神经酰胺水平升高,通过抑制PLD活力,诱导肾脏系膜细胞凋亡最终导致肾脏细胞的损害。

Objective To study the mechanism of ceramide-induced cellular signal transduction and its effect on renal injury. Methods Sixty male Sprague-Dawley rats were randomly assigned to four groups （n=15）： control group, sham operation group, experiment group one and experiment group two. Except the control and the sham operation group, the other two experiment groups underwent the common bile duct bound to form the rat model of obstructive jaundice. The renal glomerular mesangial cells （GMC） were cultured primarily. After GMC were given different stimulating factors, we performed the measurement of the PLD activity and the GMC apoptosis analyzed by flow cytometry. Results The biological activity of PC-PLD was significantly decreased by TNF-α and ceramide （P〈0.05）. On the other hand, the GMC apoptosis was induced by TNF-α and ceramide （P〈0.05）. However, other opposite results were obtained in addition of ceramide inhibitor to GMC. Conclusion TNF-α can induce the ceramide increase in GMC, which is significant to effect on the kidney injury of the rats with obstructive jaundice, by the means of inducing GMC apoptosis and decreasing the activity of PC-PLD.